Enzymatic amination

Abstract

A method comprising the steps (a) contacting a hydrocarbon comprising a hydroxyl group with a biological agent having oxygen-dependent and cofactor-dependent carbohydrate oxidase activity in the presence of oxygen and carbohydrate oxidase cofactor, and (b) contacting the hydrocarbon produced in step a) with a biological agent having transaminase activity and a biological agent having cofactor-dependent amino acid dehydrogenase activity in the presence of amino acid dehydrogenase cofactor and the substrate amino acid of the amino acid dehydrogenase.

1 . A method, comprising:
 (a) contacting a hydrocarbon comprising a hydroxyl group with a first polypeptide having an oxygen-dependent and cofactor-dependent carbohydrate oxidase activity or a whole-cell biocatalyst comprising the first polypeptide in the presence of oxygen and carbohydrate oxidase cofactor; and   (b) contacting a hydrocarbon produced in the contacting (a) with a second polypeptide having a transaminase activity or a whole-cell biocatalyst comprising the second polypeptide and a third polypeptide having a cofactor-dependent amino acid dehydrogenase activity or a whole-cell biocatalyst comprising the third polypeptide in the presence of an amino acid dehydrogenase cofactor and a substrate amino acid for an amino acid dehydrogenase,   wherein the hydrocarbon comprising a hydroxyl group comprises a 5 or 6 membered ring carrying at least one substituent of —(CH 2 ) x —OH, where x is 0 to 4, and   the first polypeptide is selected from the group consisting of an M1 variant of galactose oxidase from  Fusarium  NRRL 2903, pyranose oxidase from  Phanerochaete chrysosporium,  hexose oxidase from  Chondrus crispus  and a homologue thereof.   
     
     
         2 . The method according to  claim 1 ,
 wherein the hydrocarbon comprising a hydroxyl group has formula (I) or (II):   
       
         
           
           
               
               
           
         
         wherein one or two of A, B, C, D, E and F are atoms each and independently selected from the group consisting of N, S and O and others are C, 
         R 3  is selected from the group consisting of H, halogen, substituted or unsubstituted alkyl, alkenyl, alkynyl, —(CH 2 ) x —CR 4 O, —(CH 2 ) x —NH 2 , —(CH 2 ) x —NO 2 , —(CH 2 ) x —O—R 4 , —(CH 2 ) x —CH 2 SH, and —(CH 2 ) x —COOR 4 , 
         where x is 0 to 20, and 
         R 4  is selected from the group consisting of H, a substituted or unsubstituted alkyl, a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted aryl, and a substituted or unsubstituted heteroaryl. 
       
     
     
         3 . The method according to  claim 2 ,
 wherein one or two of A, B, C, D, E and F are atoms each and independently selected from the group consisting of N and O, and others are C,   R 3  is selected from the group consisting of —(CH 2 ) x —CR 4 O, —(CH 2 ) x —NH 2 , —(CH 2 ) x —O—R 4 , and —(CH 2 ) x —COOR 4 ,   where x is 0 to 4,   and R 4  is selected from the group consisting of H 2  and an alkyl comprising 1 to 4 carbon atoms.   
     
     
         4 . The method according to  claim 3 ,
 wherein the hydrocarbon comprising a hydroxyl group is a cycloalkanol.   
     
     
         5 . The method according to  claim 1 ,
 wherein the amino acid dehydrogenase is alanine dehydrogenase.   
     
     
         6 . The method according to  claim 1 ,
 wherein the transaminase is selected from the group consisting of ω-transaminase from  Vibrio fluvialis,  ω-transaminase from  Paracoccus denitrificans,  and a homologue thereof.   
     
     
         7 . The method according to  claim 1 ,
 wherein the contacting (b) is carried out in the presence of a fourth polypeptide having a formate dehydrogenase activity or a whole-cell biocatalyst comprising the fourth polypeptide.   
     
     
         8 . The method according to  claim 1 ,
 wherein the contacting (b) is carried out in the presence of a fifth polypeptide having a glucose dehydrogenase activity or a whole-cell biocatalyst comprising the fifth polypeptide.   
     
     
         9 . (canceled) 
     
     
         10 . The method according to  claim 7 ,
 wherein a redox factor produced by the amino acid dehydrogenase activity of the third polypeptide is consumed by the formate dehydrogenase activity of the fourth polypeptide.   
     
     
         11 . The method according to  claim 1 ,
 wherein the contacting (a) is carried out in the presence of a compound having an H 2 O 2 -degrading activity.   
     
     
         12 . The method according to  claim 11 ,
 wherein the compound is a seventh polypeptide comprising catalase and horse radish peroxidase/ABTS, and a homologue thereof or a whole-cell biocatalyst comprising the seventh polypeptide.   
     
     
         13 . The method according to  claim 1 , wherein the contacting (a) and the contacting (b) are carried out simultaneously in one reaction mixture. 
     
     
         14 . The method according to  claim 12 ,
 wherein the contacting (a) and the contacting (b) are carried out in a reaction mixture, and the second polypeptide or the whole-cell biocatalyst comprising the second polypeptide is added to the reaction mixture after the first polypeptide or the whole-cell biocatalyst comprising the first polypeptide is added to the reaction mixture.   
     
     
         15 . The method according to  claim 1 ,
 wherein a level of oxygen pressure during the contacting (a) and the contacting (b) is from 2 to 7 bar.   
     
     
         16 . The method according to  claim 1 ,
 wherein at least one of the first, second, and third polypeptides is associated with a viable cell.   
     
     
         17 . A method for transaminating a hydrocarbon comprising a hydroxyl group, the method comprising:
 transaminating the hydrocarbon with use-of a mixture comprising
 a first polypeptide having an oxygen-dependent and cofactor-dependent carbohydrate oxidase activity or a whole-cell biocatalyst comprising the first polypeptide, 
 a second polypeptide having a transaminase activity or a whole-cell biocatalyst comprising the second polypeptide, 
 a third polypeptide having an amino acid dehydrogenase activity or a whole-cell biocatalyst comprising the third polypeptide, 
   oxygen,   a substrate amino acid for an amino acid dehydrogenase,   carbohydrate oxidase cofactor, and   an amino acid dehydrogenase cofactor,   wherein the hydrocarbon comprising a hydroxyl group comprises a 5 or 6 membered ring carrying at least one substituent of —(CH 2 ) x —OH,   where x is 0 to 4, and   the first polypeptide is selected from the group consisting of an M1 variant of galactose oxidase from  Fusarium  NRRL 2903, pyranose oxidase from  Phanerochaete chrysosporium,  hexose oxidase from  Chondrus crispus,  and a homologue thereof.   
     
     
         18 . An aqueous reaction mixture, comprising:
 a hydrocarbon comprising a hydroxyl group;   a first polypeptide having an oxygen-dependent and cofactor-dependent carbohydrate oxidase activity or a whole-cell biocatalyst comprising the first polypeptide;   oxygen;   a carbohydrate oxidase cofactor;   a second polypeptide having a transaminase activity or a whole-cell biocatalyst comprising the second polypeptide;   a third polypeptide having an amino acid dehydrogenase activity or a whole-cell biocatalyst comprising the third polypeptide;   an amino acid dehydrogenase cofactor; and   a substrate amino acid for an amino acid dehydrogenase,   wherein at least one of the first, second, and third polypeptides and the whole-cell biocatalysts comprising the first, second and third polypeptides comprises, respectively, a heterologous polypeptide having a respectively activity,   the hydrocarbon comprising a hydroxyl group comprises a 5 or 6 membered ring having at least one substituent of —(CH 2 ) x —OH, where x is 0 to 4, and   the first polypeptide is selected from the group consisting of an M1 variant of galactose oxidase from  Fusarium  NRRL 2903, pyranose oxidase from  Phanerochaete chrysosporium,  hexose oxidase from  Chondrus crispus,  and homologues a homologue thereof.